The need to develop an effective method for hepatocellular carcinoma (HCC) diagnosis is urgent. even with identification of cancer DNA biomarkers, The low concentration and fragmentation of cfDNA presents a formidable challenge to the translation of DNA markers to clinic. JBS Science Inc. has developed innovative solutions to these challenges by: (1) development of sensitive short-amplicon PCR assays, suitable for cfDNA, and (2) use of urine as the source for cfDNA that provides a high volume. JBS innovations are suitable for developing cfDNA-based assays in general. HCC is our first application. We have developed the cfDNA-based assays for three validated HCC markers, TP53, CTNNB1, and TERT mutations. The Phase I is to standardize assay parameters and specimen selection (urine or plasma), culminating in a standard operating procedure (SOP) with an analytic sensitivity of 10 copies with a specificity of 1:1000, a linearity of 4 logs, and a co-efficiency of variation (reproducibility) of 0.99-1.0, and to demonstrate at least 80% sensitivity for HCC. In Phase II, we will deliver a CLIA-dertified cfDNA-based test that can be finished in one day for HCC diagnosis, so that 30% more HCC can be detected early to improve prognosis.